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Optimization of protein crystallization: The opticryst project

机译：蛋白质结晶的优化：光学项目

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Protein crystallization has gained a new strategic and commercial relevance in the postgenomic era due to its pivotal role in structural genomics. Producing high quality crystals has always been a bottleneck to efficient structure determination, and this problem is becoming increasingly acute. This is especially true for challenging, therapeutically important proteins that typically do not form suitable crystals. The OptiCryst consortium has focused on relieving this bottleneck by making a concerted effort to improve the crystallization techniques usually employed, designing new crystallization tools, and applying such developments to the optimization of target protein crystals. In particular, the focus has been on the novel application of dual polarization interferometry (DPI) to detect suitable nucleation; the application of in situ dynamic light scattering (DLS) to monitor and analyze the process of crystallization; the use of UV-fluorescence to differentiate protein crystals from sa the design of novel nucleants and seeding technologies; and the development of kits for capillary counterdiffusion and crystal growth in gels. The consortium collectively handled 60 new target proteins that had not been crystallized previously. From these, we generated 39 crystals with improved diffraction properties. Fourteen of these 39 were only obtainable using OptiCryst methods. For the remaining 25, OptiCryst methods were used in combination with standard crystallization techniques. Eighteen structures have already been solved (30% success rate), with several more in the pipeline.
机译：由于蛋白质结晶在结构基因组学中的关键作用，因此在后基因组学时代已获得了新的战略和商业意义。生产高质量的晶体一直是有效确定结构的瓶颈，这个问题变得越来越严重。对于通常无法形成合适晶体的具有挑战性的，治疗上重要的蛋白质而言，尤其如此。 OptiCryst联盟致力于通过共同努力改善通常采用的结晶技术，设计新的结晶工具并将此类开发成果应用于目标蛋白质晶体的优化来消除这一瓶颈。尤其是，重点放在了双极化干涉仪（DPI）在检测合适的成核作用上的新应用上。原位动态光散射（DLS）在监测和分析结晶过程中的应用;利用紫外线荧光将蛋白质晶体与盐区分开;设计新型成核剂和播种技术;以及用于凝胶中的毛细管反扩散和晶体生长的试剂盒的开发。该财团共同处理了60种新的靶蛋白，这些蛋白先前尚未结晶。由此，我们生成了39个具有改进衍射特性的晶体。这39个中只有14个可以使用OptiCryst方法获得。对于其余的25种，将OptiCryst方法与标准结晶技术结合使用。已经解决了18个结构（成功率30％），还有几个正在开发中。

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